Dinsmore J H, Sloboda R D
Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire 03755.
Cell. 1989 Apr 7;57(1):127-34. doi: 10.1016/0092-8674(89)90178-5.
Previously, we described a 62 kd protein that is a component of the isolated sea urchin mitotic apparatus. This protein is a substrate for an endogenous, calcium/calmodulin-dependent protein kinase also associated with the mitotic apparatus. Phosphorylation of the 62 kd protein directly correlates with the depolymerization of microtubules in isolated mitotic apparatuses. Here we report a test of the function of the 62 kd protein in vivo. Double labeling studies using a monoclonal antibody to tubulin and an affinity purified antibody specific for the 62 kd protein reveal that the 62 kd protein co-localizes with mitotic apparatus microtubules. When affinity purified antibodies to the 62 kd protein were microinjected into dividing sea urchin embryos, mitosis was blocked in a stage-specific manner. The results are discussed with respect to the role of the 62 kd protein in the metaphase-anaphase transition.
此前,我们描述了一种62kd的蛋白质,它是分离出的海胆有丝分裂器的一个组成部分。这种蛋白质是一种内源性钙/钙调蛋白依赖性蛋白激酶的底物,该激酶也与有丝分裂器相关。62kd蛋白质的磷酸化与分离出的有丝分裂器中微管的解聚直接相关。在此,我们报告了对62kd蛋白质在体内功能的一项测试。使用抗微管蛋白单克隆抗体和对62kd蛋白质特异的亲和纯化抗体进行的双重标记研究表明,62kd蛋白质与有丝分裂器微管共定位。当将针对62kd蛋白质的亲和纯化抗体显微注射到正在分裂的海胆胚胎中时,有丝分裂以阶段特异性方式被阻断。我们结合62kd蛋白质在中期 - 后期转变中的作用对结果进行了讨论。